A probiotic and prebiotic composition and methods of preparation, uses thereof

Abstract

This invention belongs to the food or pharmaceutical field, specifically relating to a probiotic and prebiotic composition, its preparation process, and its application. The composition includes probiotics, resistant dextrin, xylooligosaccharides, fructooligosaccharides, stachyose, erythritol, and cranberry powder, prepared using a fluidized bed spray granulation process. As an adjuvant to antibiotics, this composition has been verified in animal and clinical trials to synergistically treat upper / lower respiratory tract infections and urinary tract infections, shorten symptom duration, accelerate recovery from the primary disease, maintain intestinal flora diversity, increase the abundance of beneficial bacteria such as Bifidobacteria, reduce the incidence of adverse reactions, and alleviate antibiotic-associated diarrhea. This invention provides a microecological restoration solution for antibiotic treatment, possessing clear clinical value and technological feasibility.

Description

Technical Field

[0001] This invention relates to the food or pharmaceutical field, and in particular to a composition, preparation process and application of a probiotic and prebiotic composition. Background Technology

[0002] In recent years, the problem of antibiotic resistance caused by overuse has become increasingly serious worldwide. According to a 2024 report by the World Health Organization (WHO), antibiotic resistance directly causes more than 1.27 million deaths annually, with an indirect linked death toll of 4.95 million. While β-lactam antibiotics (such as amoxicillin-clavulanate potassium) are widely used to treat respiratory and urinary tract infections, their overuse not only accelerates the evolution of resistant strains but also significantly disrupts the balance of gut microbiota, causing side effects such as diarrhea and constipation. A survey of Chinese medical institutions showed that approximately 30% of antibiotic users experience symptoms related to gut microbiota dysbiosis, with children and immunocompromised individuals at higher risk. Patients with related intestinal complications require additional treatment, increasing the burden on the healthcare system.

[0003] Traditional strategies such as antibiotic rotation and the development of new antibiotics have limited effectiveness. The WHO points out that only four investigational antibiotics globally are effective against drug-resistant bacteria between 2017 and 2023, and R&D investment continues to shrink due to low commercial returns. Against this backdrop, developing adjuvant formulations that can both enhance antibiotic efficacy and protect the gut microbiota has become an important direction.

[0004] To alleviate the side effects of antibiotics, although the concept of synergistic use of probiotics and prebiotics (synbiotics) has been widely recognized, there are significant drawbacks: such as unreasonable strain selection and ratios, with most formulations containing only 2-3 types of Lactobacillus or Bifidobacterium, lacking the ability to target and repair antibiotic-induced dysbiosis; the prebiotic combination is too singular, relying on traditional ingredients such as fructooligosaccharides or inulin, neglecting the synergistic effect of resistant dextrin and xylooligosaccharides; the lack of synergistic components and the failure to integrate prebiotics, making it impossible to continuously promote probiotic colonization; process defects lead to low viability, with conventional tableting or freeze-drying processes resulting in a probiotic survival rate of less than 10% in gastric acid, and uneven particle size affecting intestinal colonization efficiency; and a lack of clinical suitability: few products are designed for combination therapy with specific antibiotics (such as Junerqing), failing to achieve precise synergy between gut microbiota regulation and anti-infective treatment.

[0005] Currently, innovative solutions are urgently needed to address antibiotic-induced gut microbiota imbalance. This composition, through multi-strain synergy, synbiotic enhancement, and process optimization, provides a highly efficient and stable antibiotic adjuvant for clinical use, demonstrating significant research value and application prospects. Summary of the Invention

[0006] Although there are various probiotic compositions in the existing technology, problems such as unreasonable strain selection and ratio, monotonous prebiotic combination, lack of synergistic components, low viability due to process defects, and lack of clinical suitability have not been properly solved. The present invention overcomes the shortcomings of the existing technology and provides a highly efficient and stable antibiotic adjuvant preparation for clinical use through multi-strain synergy, synbiotic enhancement and process optimization.

[0007] Specifically, the technical solution and technical effects of the present invention are as follows:

[0008] One of the objectives of this invention is to provide a probiotic and prebiotic composition, wherein the composition comprises probiotics, resistant dextrin, xylooligosaccharides, fructooligosaccharides, stachyose, erythritol and vitamin C.

[0009] In some embodiments, the probiotics include Bifidobacterium animalis subsp. lactis BLa80, Pediococcus lactis CCFM7902, Lactobacillus acidophilus LA85, Lactobacillus plantarum N13, Lactobacillus paracasei LC86, Bifidobacterium bifidum BBi32, Lactobacillus rhamnosus LRa05, and Lactobacillus casei LC89.

[0010] In a preferred embodiment, the strains are expressed as follows by weight:

[0011] Bifidobacterium animalis subsp. lactis BLa80 10%-20%

[0012] Pediococcus acidilactici CCFM7902 5%-15%

[0013] Lactobacillus acidophilus LA85 5%-15%

[0014] Lactobacillus plantarum N13 10%-20%

[0015] Lactobacillus paracasei LC86 10%-20%

[0016] Bifidobacterium bifidum BBi32 5%-15%

[0017] Lactobacillus rhamnosus LRa05 10%-20%

[0018] Lactobacillus casei LC89 5%-15%.

[0019] A second objective of this invention is to provide a weight ratio of a probiotic and prebiotic composition, and to determine the optimal ratio of probiotics and prebiotics, wherein the composition comprises the following components:

[0020] Probiotics 1-50 servings by weight

[0021] 5-100 parts by weight of resistant dextrin

[0022] 5-100 parts by weight of xylooligosaccharides

[0023] 1-50 parts by weight of fructooligosaccharides

[0024] Stachyose 1-50 parts by weight

[0025] 1-50 parts by weight of erythritol

[0026] Cranberry powder, 1-50 parts by weight.

[0027] In a preferred embodiment, the composition comprises the following components:

[0028] 5-20 servings of probiotics

[0029] 10-50 parts by weight of resistant dextrin

[0030] 10-50 parts by weight of xylooligosaccharides

[0031] 3-10 parts by weight of fructooligosaccharides

[0032] 1-5 parts by weight of stachyose

[0033] 5-10 parts by weight of erythritol

[0034] 5-10 parts by weight of cranberry powder.

[0035] A third objective of this invention is to provide a probiotic and prebiotic composition containing vitamin C.

[0036] In some embodiments, the vitamin C comprises natural plant extracts and synthetic vitamin C and its derivatives.

[0037] In a more preferred embodiment, the vitamin C is natural vitamin C, and further, the vitamin C is cranberry powder.

[0038] The fourth objective of this invention is to screen erythritol as a binder for granulation and to determine the optimal process. In formulation process development, the screening of binders and the optimization of granulation parameters are crucial to ensuring the stability and functionality of probiotic compositions.

[0039] First, through orthogonal experiments, the granulation performance of common adhesives on the market, such as erythritol, dextrin, and microcrystalline cellulose, was compared, and erythritol was determined to be the optimal adhesive for this solution.

[0040] Secondly, through orthogonal experiments: air inlet temperature (50 / 60 / 70℃) × atomization pressure (0.1 / 0.2 / 0.3MPa) × fluidization velocity (1.5 / 2.0 / 2.5m / s), the preparation process of the present invention was determined to be as follows: (1) erythritol was prepared into an adhesive solution; (2) resistant dextrin and xylooligosaccharide were mixed, sprayed into the adhesive of step (1) for granulation, and after sieving, it was mixed with stachyose, fructooligosaccharide, cranberry powder and probiotic powder to obtain the final product.

[0041] In a more preferred embodiment, the preparation process of the present invention is as follows: (1) Erythritol is prepared into a binder solution with a concentration of 10-50% (w / v); (2) Resistant dextrin and xylooligosaccharide are mixed and sprayed into the binder of step (1) for granulation. The granulation particle size is controlled to be 0.3-1.0 mm. After sieving, it is mixed with stachyose, fructooligosaccharide, cranberry powder and probiotic powder to obtain the final product. Further, the air inlet temperature of the granulation process is 50-70℃, the atomization pressure is 0.1-0.3 MPa, and the material fluidization velocity is 1.5-3.0 m / s.

[0042] The fifth objective of this invention is to provide an application of a probiotic and prebiotic composition in the preparation of antibiotic adjuvants.

[0043] In some embodiments, the probiotic and prebiotic composition is used to alleviate antibiotic-induced gut microbiota imbalance, diarrhea, or constipation, while regulating the gut microbiota.

[0044] In some embodiments, the composition is used in combination with an antibiotic preparation for the treatment of related disease infections.

[0045] In a preferred embodiment, the related disease infections include upper respiratory tract infections, lower respiratory tract infections, urinary tract infections, skin and soft tissue infections, and other infections.

[0046] Compared with the prior art, the present invention achieves the following significant technical effects:

[0047] (1) The probiotic and prebiotic composition provided by the present invention has a regulatory effect on the intestinal flora of mice with antibiotic-induced diarrhea, which is beneficial to the growth of mouse body weight, reduces the rate of loose stools in mice, increases the level of serum immunoglobulin IgA and IgG in mice, can significantly improve the low immune function of mice caused by AAD, has a significant intestinal flora regulation effect on AAD mice, and can alleviate the symptoms of antibiotic-related diarrhea.

[0048] (2) In the experiment of treating upper respiratory tract infection with the combination of probiotic and prebiotic composition of the present invention and amoxicillin clavulanate potassium tablets, the body weight of mice increased, the lung index of mice in each administration group decreased significantly, and the levels of TNF-α and IL-8 decreased. The combined administration can suppress the inflammatory response of mice and improve lung tissue damage in mice.

[0049] (3) The combination of probiotics and prebiotics in this invention can reduce the organ index of rats with acute pyelonephritis, reduce the levels of serum creatinine, urine creatinine and serum IL-1β and CXCL-2, and significantly increase the levels of urine SIgA and serum IL-10, suggesting that the combined use of drugs can reduce kidney damage, inhibit the inflammatory response in rats, and enhance the local immune function of the rat urethra.

[0050] (4) Through trial administration experiments, the probiotic composition of the present invention can reduce the incidence of adverse reactions, shorten the duration of symptoms, accelerate the recovery of the primary disease, maintain the diversity of intestinal flora, increase the abundance of beneficial bacteria such as Bifidobacteria, and relieve inflammation, providing an effective solution for the precise intervention of antibiotic-related intestinal side effects. Attached Figure Description

[0051] Figure 1 The effect of administering a combination of probiotics and prebiotics on serum immunoglobulin IgA levels in mice (compared to the control group). * P <0.05, ** P <0.01; compared with the model group # P <0.05, ## P <0.01).

[0052] Figure 2 The effect of administering a combination of probiotics and prebiotics on serum immunoglobulin IgG levels in mice (compared to the control group). * P <0.05, ** P <0.01; compared with the model group # P <0.05, ## P <0.01).

[0053] Figure 3 Phylogenetic Species Analysis: Stacked Bar Chart

[0054] Figure 4 Genus-level species analysis bar stacking diagram Specific Implementation

[0055] Example 1: A probiotic and prebiotic composition and its preparation method

[0056]

[0057] Preparation process: (1) Prepare a 20% (w / v) binder solution of erythritol; (2) Mix resistant dextrin and xylooligosaccharide, spray and add the binder solution from step (1) to granulate, control the particle size to 0.7 mm, the inlet air temperature to 60 °C, the atomization pressure to 0.2 MPa, and the material fluidization velocity to 2.0 m / s. After sieving, mix with stachyose, fructooligosaccharide, cranberry powder and probiotic powder to obtain the final product.

[0058] Example 2: A probiotic and prebiotic composition and its preparation method

[0059]

[0060] Preparation process: (1) Prepare a 50% (w / v) binder solution of erythritol; (2) Mix resistant dextrin and xylooligosaccharide, spray and add the binder solution from step (1) to granulate, control the particle size to 0.3 mm, the inlet air temperature to 70 °C, the atomization pressure to 0.1 MPa, and the material fluidization velocity to 1.5 m / s. After sieving, mix with stachyose, fructooligosaccharide, cranberry powder and probiotic powder to obtain the final product.

[0061] Example 3: A probiotic and prebiotic composition and its preparation method

[0062]

[0063] Preparation process: (1) Prepare a 30% (w / v) binder solution of erythritol; (2) Mix resistant dextrin and xylooligosaccharide, spray and add the binder solution from step (1) to granulate, control the particle size to 0.8 mm, the inlet air temperature to 50 °C, the atomization pressure to 0.3 MPa, and the material fluidization velocity to 2.0 m / s. After sieving, mix with stachyose, fructooligosaccharide, cranberry powder and probiotic powder to obtain the final product.

[0064] Example 4: A probiotic and prebiotic composition and its preparation method

[0065]

[0066] Preparation process: (1) Prepare a 40% (w / v) binder solution of erythritol; (2) Mix resistant dextrin and xylooligosaccharides, spray and add the binder solution from step (1) to granulate, control the particle size to 0.5 mm, the inlet air temperature to 65 °C, the atomization pressure to 0.25 MPa, and the material fluidization velocity to 2.5 m / s. After sieving, mix with stachyose, fructooligosaccharides, cranberry powder and probiotic powder to obtain the final product.

[0067] Example 5: A probiotic and prebiotic composition and its preparation method

[0068]

[0069] Preparation process: (1) Prepare an adhesive solution with a concentration of 10-50% (w / v) of erythritol; (2) Mix resistant dextrin and xylooligosaccharide, spray and add the adhesive solution from step (1) to granulate, control the particle size to 0.6 mm, the inlet air temperature to 55 ℃, the atomization pressure to 0.15 MPa, and the material fluidization velocity to 3.0 m / s. After sieving, mix with stachyose, fructooligosaccharide, cranberry powder and probiotic powder to obtain the final product.

[0070] Comparative Example 1: A probiotic and prebiotic composition and its preparation method

[0071]

[0072] Preparation process: (1) Prepare a 20% (w / v) binder solution of erythritol; (2) Mix resistant dextrin and xylooligosaccharide, spray and add the binder solution from step (1) to granulate, control the particle size to 0.7 mm, the inlet air temperature to 60 °C, the atomization pressure to 0.2 MPa, and the material fluidization velocity to 2.0 m / s. After sieving, mix with stachyose, fructooligosaccharide, cranberry powder and probiotic powder to obtain the final product.

[0073] Comparative Example 2: A probiotic and prebiotic composition and its preparation method

[0074]

[0075] Preparation process: (1) Prepare an adhesive solution with a concentration of 10-50% (w / v) by erythritol; (2) Mix resistant dextrin and xylooligosaccharides, spray them into the adhesive solution from step (1) for granulation, control the particle size to be 0.3-1.0 mm, the inlet air temperature to be 50-70℃, the atomization pressure to be 0.1-0.3 MPa, and the material fluidization velocity to be 1.5-3.0 m / s. After sieving, mix with stachyose, fructooligosaccharides, cranberry powder and probiotic powder to obtain the final product.

[0076] Comparative Example 3: A probiotic and prebiotic composition and its preparation method

[0077]

[0078] Preparation process: (1) Prepare an adhesive solution with a concentration of 10-50% (w / v) of erythritol; (2) Spray resistant dextrin into the adhesive solution of step (1) for granulation, with the particle size controlled at 0.3-1.0 mm, the inlet air temperature at 50-70℃, the atomization pressure at 0.1-0.3 MPa, and the material fluidization velocity at 1.5-3.0 m / s. After sieving, mix with inulin, cranberry powder and probiotic powder to obtain the final product.

[0079] Comparative Example 4: A probiotic and prebiotic composition and its preparation method

[0080]

[0081] Preparation process: (1) Prepare an adhesive solution with a concentration of 10-50% (w / v) by erythritol; (2) Mix resistant dextrin and xylooligosaccharides, spray them into the adhesive solution from step (1) for granulation, control the particle size to be 0.3-1.0 mm, the inlet air temperature to be 50-70℃, the atomization pressure to be 0.1-0.3 MPa, and the material fluidization velocity to be 1.5-3.0 m / s. After sieving, mix with maltodextrin, fructooligosaccharides, cranberry powder and probiotic powder to obtain the final product.

[0082] Comparative Example 5: A probiotic and prebiotic composition and its preparation method

[0083]

[0084] Preparation process: (1) Prepare an adhesive solution with a concentration of 10-50% (w / v) of erythritol; (2) Spray resistant dextrin into the adhesive solution of step (1) for granulation, with the particle size controlled at 0.3-1.0 mm, the inlet air temperature at 50-70℃, the atomization pressure at 0.1-0.3 MPa, and the material fluidization velocity at 1.5-3.0 m / s. After sieving, mix with fructooligosaccharides, cranberry powder, and probiotic powder to obtain the final product.

[0085] Comparative Example 6: A probiotic and prebiotic composition and its preparation method

[0086]

[0087] Preparation process: (1) Prepare an adhesive solution with a concentration of 10-50% (w / v) by erythritol; (2) Mix resistant dextrin and xylooligosaccharides, spray them into the adhesive solution from step (1) for granulation, control the particle size to be 0.3-1.0 mm, the inlet air temperature to be 50-70℃, the atomization pressure to be 0.1-0.3 MPa, and the material fluidization velocity to be 1.5-3.0 m / s. After sieving, mix with stachyose, fructooligosaccharides, cranberry powder and probiotic powder to obtain the final product.

[0088] Comparative Example 7: A probiotic and prebiotic composition and its preparation method

[0089]

[0090] Preparation process: (1) Prepare an adhesive solution with a concentration of 10-50% (w / v) by erythritol; (2) Mix xylooligosaccharides and fructooligosaccharides, spray them into the adhesive solution from step (1) for granulation, control the particle size to be 0.3-1.0 mm, the inlet air temperature to be 50-70℃, the atomization pressure to be 0.1-0.3 MPa, and the material fluidization velocity to be 1.5-3.0 m / s. After sieving, mix with stachyose, cranberry powder and probiotic powder to obtain the final product.

[0091] Comparative Example 8: A probiotic and prebiotic composition and its preparation method

[0092]

[0093] Preparation process: (1) Prepare a binder solution with a concentration of 10-50% (w / v) by microcrystalline cellulose; (2) Mix resistant dextrin and xylooligosaccharides, spray them into the binder solution from step (1) and granulate them. The particle size is controlled at 0.3-1.0 mm, the air inlet temperature is 50-70℃, the atomization pressure is 0.1-0.3 MPa, and the material fluidization velocity is 1.5-3.0 m / s. After sieving, mix with stachyose, fructooligosaccharides, cranberry powder and probiotic powder to obtain the final product.

[0094] Comparative Example 9: A probiotic and prebiotic composition and its preparation method

[0095]

[0096] Preparation process: (1) Prepare a binder solution with microcrystalline cellulose at a concentration of 10 - 50% (w / v); (2) Mix resistant dextrin and xylooligosaccharide, and spray and add them to the binder in step (1) for granulation. Control the granule size to be 0.3 - 1.0 mm, the inlet air temperature to be 50 - 70°C, the atomization pressure to be 0.1 - 0.3 MPa, and the material fluidization speed to be 1.5 - 3.0 m / s. After sieving, mix with stachyose, fructooligosaccharide, L-ascorbic acid, and probiotic powder to obtain the product.

[0097] Comparative Example 10: A probiotic and prebiotic composition and its preparation method

[0098]

[0099] Preparation process: (1) Prepare a binder solution with erythritol at a concentration of 60% (w / v); (2) Mix resistant dextrin and xylooligosaccharide, and spray and add them to the binder in step (1) for granulation. Control the granule size to be 1.5 mm, the inlet air temperature to be 80°C, the atomization pressure to be 0.4 MPa, and the material fluidization speed to be 1.0 m / s. After sieving, mix with stachyose, fructooligosaccharide, cranberry powder, and probiotic powder to obtain the product.

[0100] Verification test example

[0101] I. Regulatory effect of the probiotic and prebiotic composition of the present invention on the intestinal flora of mice with antibiotic-induced diarrhea model

[0102] 1 Materials and methods

[0103] 1.1 Materials

[0104] 1.1.1 Experimental drugs

[0105] Probiotic and prebiotic composition: Example 1, Example 2, Comparative Example 1, Comparative Example 3, Comparative Example 6, Comparative Example 7. Ampicillin sodium for injection (Shanghai Yuanye Bio-Technology Co., Ltd.); Bifico (Shanghai Shangyao Xinyi Pharmaceutical Factory Co., Ltd.).

[0106] 1.1.2 Experimental animals

[0107] 48 SPF-grade male BALB / c mice, 6 - 8 weeks old, weighing 18 - 20 g, provided by Shandong Pengyue Experimental Animal Technology Co., Ltd., animal certificate number: SCXK(Lu) 2022 0006. The experimental animals were housed in a barrier environment animal room, with free access to water and food during the period, the temperature was controlled at 20 - 23°C, and there was a 12-hour light-dark cycle.

[0108] 1.2 Experimental methods

[0109] 1.2.1 Grouping and model construction

[0110] After 1 week of acclimatization, BALB / c mice were randomly divided into 9 groups of 8 mice each: blank group, model group, positive group (Bifidobacterium), Example 1 group, Example 2 group, comparative example 1 group, comparative example 3 group, comparative example 6 group, and comparative example 7 group.

[0111] Mice in the model group, positive group, and probiotic group were administered ampicillin (22.4 g / kg) by gavage twice daily, with each dose being 11.2 g / kg. Mice in the control group were administered an equal volume of physiological saline by gavage at the same time daily for 3 consecutive days. The criteria for successful model establishment were: mice curled up, reduced activity, lethargy, soiled perianal area, and obvious stains on the buttocks; or statistically significant differences in diarrhea rate and diarrhea index between the model group and the control group.

[0112] After successful model construction, probiotic preparations were administered. The control group and model group were given saline by gavage, while the positive control group was given Bifidobacterium (8 g / kg, with a live bacteria count of no less than 1×10⁻⁶ per mouse) by gavage. 7 Mice in the CFU, probiotic, and prebiotic groups were each given 5 × 10⁻⁶ CFU of probiotic preparation. 6 CFU, for 14 consecutive days.

[0113] 1.2.2 Effects of probiotic and prebiotic combination on mouse body weight

[0114] On the first day of the experiment, which was the first day of establishing the diarrhea model, at 10:00 AM, the initial weight of each group of mice was recorded. Then, the mice were given ampicillin. At 6:00 PM every other day, the weight of each group of mice was weighed and recorded.

[0115] 1.2.3 Effects of probiotics and prebiotic preparations on the rate of loose stools in mice

[0116] After modeling, on the 3rd day of the experiment and 14 days after the administration of probiotics, on the 17th day of the experiment, the number of loose stools in each group of mice was observed and recorded, and the loose stool rate of mice was calculated: the ratio of the number of loose stools excreted by each animal to the total number of stools.

[0117] 1.2.4 Effects of probiotic and prebiotic combination on serum immunoglobulin IgA and IgG levels in mice

[0118] One hour after the last administration of the probiotic preparation, blood was collected from the orbital sinus of mice in each group. The samples were centrifuged at 3000 r / min for 10 min, and the serum was aliquoted and frozen at -80℃ for later testing. Serum IgA and IgG levels were detected using an ELISA kit.

[0119] 1.2.5 Effects of probiotic and prebiotic combinations on the gut microbiota of mice

[0120] After modeling and 30 min after the last administration of probiotic preparation, mouse feces were aseptically collected. The samples were diluted 10-fold to 10⁻⁸. Appropriate dilutions were selected, and the samples were inoculated onto BBL agar, MRS agar, TSC agar, and modified GAM agar media, respectively. After incubation, the levels of Bifidobacterium, Lactobacillus, Clostridium perfringens, and Enterococcus per gram of wet feces were calculated. The results were expressed as the logarithm of colony-forming units per unit mass of feces (lg CFU / g).

[0121] 1.2.6 Statistical Analysis

[0122] SPSS 26.0 statistical software was used. Experimental data are expressed as mean ± standard deviation. The mean ± standard deviation (S) indicates that a one-way ANOVA method was used to test the significance of two or more groups of data. P A value < 0.05 indicates a significant difference; plotted using Sigmaplot14.

[0123] 2 Experimental Results

[0124] 2.1 Effects of probiotic and prebiotic combination on mouse body weight

[0125] Table 1. Effects of the probiotic and prebiotic composition of the present invention on the weight changes of mice in each group.

[0126]

[0127] Compared with the blank group * P <0.05, ** P <0.01; compared with the model group # P <0.05, ## P <0.01.

[0128] As shown in Table 1, compared with the blank group, the weight gain of mice in each group was slow during the modeling period. Compared with the model group, there was no significant difference in weight gain among the probiotic preparation groups, indicating that the model was successfully established. After 14 days of probiotic administration, compared with the blank group, there was no significant difference in weight gain in the model group, while the positive group showed a weight gain of 46.38% ( P <0.01), the weight gain in Example 1 group was 54.35% ( P <0.01), the weight gain in Example 2 group was 37.32% ( P <0.05); compared with the model group, the body weight of mice in the positive group, Example 1 and 2 groups increased by 60.96% ( P <0.01), 69.72% P <0.01), 51%P <0.01), the effect of the Example 1 group was better than that of the positive group, and the effect of the Example 2 group was comparable to that of the positive group.

[0129] 2.2 Effects of probiotics and prebiotics on the rate of loose stools in diarrheal mice

[0130] Table 2. Effects of the probiotic and prebiotic composition of the present invention on the rate of loose stools in diarrheal mice.

[0131]

[0132] Compared with the blank group * P <0.05, ** P <0.01; compared with the model group # P <0.05, ## P <0.01.

[0133] As shown in Table 2, compared with the blank group, the rate of loose stools in the other groups of mice after modeling was significantly different. P <0.01), after 14 days of probiotic administration, compared with the blank control group, the rate of loose stools in the other groups of mice was significantly different ( P <0.01), compared with the model group, the rate of loose stools in mice was significantly reduced in the positive group and the Example 1 and 2 groups, with the most significant reduction in the Example 1 group, suggesting that its effect may be better than the positive control group, while the Example 2 group had a similar effect to the positive group.

[0134] 2.3 Effects of probiotics and prebiotic preparations on serum immunoglobulin IgA and IgG levels in mice

[0135] Depend on Figure 1 , Figure 2 It can be seen that after taking probiotics and prebiotics for 14 days, compared with the blank group, the levels of IgA and IgG in the model group were significantly reduced. P <0.01), compared with the model group, the levels of IgA and IgG in the positive control group and comparative groups 1 and 2 were significantly increased ( P <0.05), the increase in serum immunoglobulin IgA and IgG levels in mice can significantly improve the weakened immune function in mice caused by AAD.

[0136] 2.4 Effects of probiotic and prebiotic combinations on the gut microbiota of mice in each group

[0137] Table 3. Changes in intestinal flora in mice before and after administration of the probiotic and prebiotic composition of the present invention.

[0138]

[0139] Note: * P <0.05, ** P <0.01, compared with 0d.

[0140] Table 3 shows the changes in gut microbiota in mice before and after probiotic administration. On day 14, compared with the control group, the number of Bifidobacterium and Lactobacillus was significantly decreased, while the number of Enterococcus and Clostridium perfringens was significantly increased in the model group, indicating an imbalance in the gut microbiota of the model mice. Compared with before administration (day 0), the gut microbiota in the model group recovered to some extent on day 14, but none reached a healthy level. In the positive group and Examples 1 and 2, Bifidobacterium, Lactobacillus, Enterococcus, and Clostridium perfringens all showed some recovery. P <0.05), with the positive group and Example 1 group showing the greatest recovery, reaching the level of the blank group. Bifidobacteria and lactobacilli are among the dominant bacteria in the human or animal gut, enhancing intestinal barrier function through multiple pathways. When the gut microbiota is imbalanced, the relative abundance of Bifidobacteria and lactobacilli decreases, while the relative abundance of enterococci and opportunistic pathogens such as Clostridium perfringens, the main pathogen of AAD, increases significantly, thereby inducing intestinal diseases. Experimental results show that the probiotic and prebiotic composition of this invention has a significant gut microbiota regulatory effect on AAD mice and can alleviate antibiotic-associated diarrhea symptoms.

[0141] II. A study on the use of probiotic and prebiotic combinations as an adjunct to amoxicillin-clavulanate potassium in the treatment of upper respiratory tract infections.

[0142] 1. Materials and Methods

[0143] 1.1 Materials

[0144] 1.1.1 Experimental Drugs

[0145] Probiotic and prebiotic composition: Examples 1, 3, 5, Comparative Example 2, Comparative Example 4, and Comparative Example 5. Amoxicillin Clavulanate Potassium Tablets (Lunan Better Pharmaceutical Co., Ltd.)

[0146] 1.1.2 Laboratory Animals and Grouping

[0147] BALB / c mice, 4-6 weeks old, weighing 14-18g, half male and half female, were randomly divided into 9 groups (n=8 per group) after 1 week of acclimatization: blank group, model group, amoxicillin-clavulanate potassium tablet group, amoxicillin-clavulanate potassium + Example 1 group, amoxicillin-clavulanate potassium + Example 3 group, amoxicillin-clavulanate potassium + Example 5 group, amoxicillin-clavulanate potassium + control Example 2 group, amoxicillin-clavulanate potassium + control Example 4 group, and amoxicillin-clavulanate potassium + control Example 5 group.

[0148] 1.2 Experimental Methods

[0149] 1.2.1 Model Construction

[0150] Administer 0.1 ml (0.5 mg) of hydrocortisone intraperitoneally every morning and 0.2 ml (0.5 mg) of cyclophosphamide intraperitoneally every afternoon, repeating this administration for 3 days. In immunosuppressed mice, 1–2 hours after the last cyclophosphamide administration, inject MRSA (methicillin-resistant Staphylococcus aureus) bacterial solution into a pediatric scalp vein needle inserted approximately 0.5 cm deep on the upper right back, about 1 cm from the base of the right ear (after disinfection). The bacterial count per mouse was 6 × 10⁻⁶. 8 CFU injection was administered to animals that died within 5 hours after injection; these were considered non-infectious deaths and were discarded. Approximately 5–12 hours after infection, the model animals exhibited lethargy, loss of appetite, sluggish response to external stimuli, a slightly arched back, and erect hairs, followed by limb paralysis (with the hind limbs often extended backward) and inability to support the body. Severe lung lesions resulted in shallow and rapid breathing, and some mice also developed diarrhea during laboratory observation.

[0151] 1.2.2 Grouping and Dosing

[0152] Control group: Uninfected with MRSA + normal saline gavage

[0153] Model group: infected with MRSA + administered via gavage with normal saline

[0154] Amoxicillin-clavulanate potassium group (AMX): MRSA + amoxicillin-clavulanate potassium tablets

[0155] Amoxicillin Clavulanate Potassium + Example 1 Group (AMX+S1): MRSA + Amoxicillin Clavulanate Potassium Tablets + Example 1 Group Probiotic and Prebiotic Composition

[0156] Amoxicillin Clavulanate Potassium + Example 3 Group (AMX+S3): MRSA + Amoxicillin Clavulanate Potassium Tablets + Example 3 Group Probiotic and Prebiotic Composition

[0157] Amoxicillin Clavulanate Potassium + Example 5 Group (AMX+S5): MRSA + Amoxicillin Clavulanate Potassium Tablets + Example 5 Group Probiotic and Prebiotic Composition

[0158] Amoxicillin Clavulanate Potassium + Comparative Example 2 (AMX+D2): MRSA + Amoxicillin Clavulanate Potassium Tablets + Comparative Example 2 Probiotic and Prebiotic Combination

[0159] Amoxicillin Clavulanate Potassium + Comparative Example 4 (AMX+D4): MRSA + Amoxicillin Clavulanate Potassium Tablets + Comparative Example Group Probiotic and Prebiotic Composition

[0160] Amoxicillin Clavulanate Potassium + Comparative Example 5 (AMX+D5): MRSA + Amoxicillin Clavulanate Potassium Tablets + Comparative Example 5 Probiotic and Prebiotic Combination

[0161] The dosage of amoxicillin-clavulanate potassium tablets is as follows: Grind the amoxicillin-clavulanate potassium tablets and dissolve them in physiological saline to a concentration of 5.35 mg / mL. Administer 0.2 mL orally three times daily. Mice in both the probiotic and prebiotic groups were given 5 × 10⁻⁶ probiotic preparations. 6 CFU was administered to the control group and the model group in equal volumes of saline for 7 days.

[0162] 1.2.3 Body weight and lung index

[0163] After administration of the drug to the mice, they were deprived of water and food the day before dissection. After weighing their feces and body weight, they were anesthetized by intraperitoneal injection of sodium pentobarbital and euthanized. Blood was collected from the heart, the entire lung was removed and the connective tissue was eliminated. The weight was determined using a 0.01 g electronic balance, and the lung index was calculated according to formula (1).

[0164] Lung Index = Lung Wet Weight / Body Weight (1)

[0165] 1.2.4 Enzyme-linked immunosorbent assay (ELISA) was used to detect the levels of inflammatory factors in mouse lung tissue.

[0166] The levels of TNF-α and IL-8 in mouse lung tissue were measured according to the steps in the ELISA kit instructions.

[0167] 1.2.5 Statistical Analysis

[0168] SPSS 26.0 statistical software was used. Experimental data are expressed as mean ± standard deviation. The mean ± standard deviation (S) indicates that a one-way ANOVA method was used to test the significance of two or more groups of data. P <0.05 indicates a significant difference.

[0169] 2 Experimental Results

[0170] 2.1 Body weight and lung index of mice in each group

[0171] Table 4. Effects of each group of drugs on body weight and lung index in mice.

[0172]

[0173] Compared with the blank group * P <0.05, ** P <0.01; compared with the model group # P <0.05, ##P <0.01.

[0174] As shown in Table 4, compared with the control group, the body weight of mice in the model group was reduced ( P <0.01); Compared with the model group, mice in the amoxicillin-clavulanate potassium and the dosage groups of Examples 1, 3, and 5 showed an increase in body weight ( P <0.01). Compared with the control group, the lung index of rats in the model group was significantly increased ( P <0.01); Compared with the model group, mice in the amoxicillin-clavulanate potassium and the dosage groups of Examples 1, 3, and 5 showed increased body weight, and the lung index of mice in each treatment group was significantly reduced ( P <0.05, P <0.01), indicating that administration of amoxicillin clavulanate potassium tablets, in combination with the probiotic and prebiotic compositions of each example group, can improve lung tissue damage in mice.

[0175] 2.2 Levels of inflammatory factors TNF-α and IL-8 in lung tissue of mice in each group

[0176] Table 5. Changes in the levels of inflammatory factors TNF-α and IL-8 in mouse lung tissue after drug administration in each group.

[0177]

[0178] Compared with the blank group * P <0.05, ** P <0.01; compared with the model group # P <0.05, ## P <0.01.

[0179] As shown in Table 5, compared with the control group, the levels of TNF-α and IL-8 in the lung tissue of the model group mice were significantly increased ( P <0.01). Compared with the model group, the amoxicillin-clavulanate potassium group and the combination of amoxicillin and clavulanate potassium groups with those in Examples 1, 3, and 5 showed significantly higher levels of TNF-α and IL-8 in the lung tissue of mice.

[0180] The water level decreased significantly ( P <0.05, P <0.01).

[0181] III. Effects of probiotic and prebiotic combinations on inflammatory factors and local immune function in rats with acute pyelonephritis

[0182] 1. Materials and Methods

[0183] 1.1 Materials

[0184] 1.1.1 Experimental animals

[0185] 60 SPF-grade SD rats, male, body weight (180 ± 20) g, provided by Lunan Pharmaceutical Group Co., Ltd., with the license number SYXK(Lu) 2023 0023. Feeding environment: temperature 18 - 28 °C, relative humidity 40% - 70%, light 12 h, alternating light and darkness.

[0186] 1.1.2 Experimental drugs

[0187] Probiotic and prebiotic composition: Example 1, Example 4, Comparative Example 8, Comparative Example 9, Comparative Example 10. Levofloxacin hydrochloride tablets, Heilongjiang Nuojie Pharmaceutical Co., Ltd.

[0188] 1.2 Experimental methods

[0189] 1.2.1 Construction of acute pyelonephritis (APN) model

[0190] After the rats were deprived of water for 18 h, they were anesthetized intraperitoneally with 2% sodium pentobarbital, fixed supine, the abdomen was depilated and disinfected, a 2-cm midline incision was made in the lower abdomen, the abdominal wall was incised layer by layer, after finding the left ureter, a No. 4 thread was passed through on both sides of the left ureter, the penis was clamped, 0.5 mL of Escherichia coli liquid was injected into the bladder, the abdominal wall was sutured layer by layer, and drinking water and diet were restored. The ureteral ligation thread was removed 24 h after the operation to open the ureter. The sham operation group was injected with 0.5 mL of normal saline. The modeling time was 3 d. The success criterion for establishing the APN model: the number of urinary white blood cells per cubic millimeter ≥ 100.

[0191] 1.2.2 Grouping and drug administration

[0192] Blank group: Sham operation + normal saline

[0193] Model group: Normal saline

[0194] Positive group: Levofloxacin hydrochloride tablets

[0195] Levofloxacin hydrochloride tablets + Example 1 group (ZYFSX + S1): Levofloxacin hydrochloride tablets + the probiotic and prebiotic composition of Example 1

[0196] Levofloxacin hydrochloride tablets + Example 4 group (ZYFSX + S4): Levofloxacin hydrochloride tablets + the probiotic and prebiotic composition of Example 4

[0197] Levofloxacin hydrochloride tablets + Comparative Example 8 group (ZYFSX + D8): Levofloxacin hydrochloride tablets + the probiotic and prebiotic composition of Example 8

[0198] Levofloxacin hydrochloride tablets + Comparative Example 9 (ZYFSX+D9): Levofloxacin hydrochloride tablets + Probiotic and prebiotic composition from Example 9

[0199] Levofloxacin hydrochloride tablets + Comparative Example 10 (ZYFSX+D10): Levofloxacin hydrochloride tablets + Probiotic and prebiotic composition from Example 10

[0200] The positive control group was given levofloxacin hydrochloride tablets 21 mg / (kg·d) plus 2 mL of normal saline by gavage. The sham surgery group and the model group were given the same volume of normal saline by gavage. The combination therapy group was given levofloxacin hydrochloride tablets 21 mg / (kg·d) plus 2 mL of a probiotic and prebiotic combination (probiotic preparation 5×10). 6 CFU was administered via gavage for 14 days.

[0201] 1.2.3 Organ Index of Rats. After intraperitoneal anesthesia, kidney and bladder tissues were harvested and measured as wet weight.

[0202] Left-to-right kidney ratio = (left kidney mass / right kidney mass) × 100%;

[0203] Kidney (bladder) index (mg / g) = Kidney (bladder) mass (mg) / Body mass (g) × 100%.

[0204] 1.2.4 Biochemical index detection.

[0205] Blood was collected from the abdominal aorta and centrifuged at 3000 r / min for 15 min. The supernatant was collected, and serum creatinine and urine creatinine were detected using the sarcosine oxidase method. IL-1β, IL-10, and CXCL-2 were detected using ELISA. Urine was collected from rats, and urine creatinine was detected using the sarcosine oxidase method. Urine SIgA was detected using ELISA.

[0206] 1.2.5 Statistical Analysis

[0207] SPSS 26.0 statistical software was used. Experimental data are expressed as mean ± standard deviation. The mean ± standard deviation (S) indicates that a one-way ANOVA method was used to test the significance of two or more groups of data. P A value < 0.05 indicates a significant difference; plotted using Sigmaplot14.

[0208] 2 Experimental Results

[0209] 2.1 Comparison of organ indices among rat groups

[0210] Table 6. Effects of drug administration on organ indices in rats.

[0211]

[0212] Compared with the blank group * P <0.05, ** P <0.01; compared with the model group # P <0.05, ## P <0.01.

[0213] As shown in Table 6, compared with the blank group, the ratio of left to right kidneys, kidney index, and bladder index of rats in the model group were significantly increased. P <0.05). Compared with the model group, the positive group and its combination therapy with Examples 1, 4, and 8 showed a significant decrease in the left-right kidney ratio, kidney index, and bladder index. P < 0.05).

[0214] 2.2 Comparison of urinary SIgA, serum creatinine, and urinary creatinine levels in rats of different groups

[0215] Table 7. Effects of each group of drugs on urinary SIgA, serum creatinine, and urinary creatinine levels in rats.

[0216]

[0217] Compared with the blank group * P <0.05, ** P <0.01; compared with the model group # P <0.05, ## P <0.01.

[0218] As shown in Table 7, compared with the blank group, the serum creatinine and urinary creatinine levels of rats in the model group were significantly increased, while the urinary SIgA level was significantly decreased. P <0.05); Compared with the model group, the serum creatinine and urinary creatinine levels of rats in the positive group and each group of the example were significantly decreased, and the urinary SIgA level was significantly increased ( P <0.05).

[0219] 2.3 Comparison of serum IL-1β, IL-10, and CXCL-2 levels in different groups of rats

[0220] Table 8. Effects of each group of drugs on serum IL-1β, IL-10, and CXCL-2 levels in rats.

[0221]

[0222] Compared with the blank group * P <0.05,** P <0.01; compared with the model group # P <0.05, ## P <0.01.

[0223] As shown in Table 8, compared with the blank group, the serum IL-1β and CXCL-2 levels in the model group rats were significantly increased, while the IL-10 level was significantly decreased. P <0.05); Compared with the model group, the serum IL-1β and CXCL-2 levels of rats in the positive group and each group of the example were significantly decreased, and the IL-10 level was significantly increased ( P <0.05).

[0224] In summary, the combination of the probiotic and prebiotic composition of the present invention with levofloxacin hydrochloride tablets can inhibit the inflammatory response in APN rats, improve local immune function, and inhibit bacterial growth.

[0225] IV. Trial of administration

[0226] 1. Objects and Methods

[0227] 1.1 General Information

[0228] Sixty patients with primary urinary tract / respiratory tract infections were recruited, with a mean age of 36.45 ± 15.75 years, including 39 males and 21 females. Baseline characteristics were balanced across groups. The trial product was a probiotic solid beverage.

[0229] Exclusion criteria: 1. History of chronic or severe diseases of the neuropsychiatric, respiratory, cardiovascular, digestive, hematologic, lymphatic, hepatic, renal, or endocrine systems. 2. Patients with active peptic ulcers. 3. Any other reason deemed unsuitable for participation by the investigator.

[0230] 1.2 Methods

[0231] 1.2.1 Grouping

[0232] Group A (n=20): Junerqing® Amoxicillin Clavulanate Potassium Dispersible Tablets + Composition of Example 1 (prepared according to the formulation of Example 1, 3g / bag, each bag contains no less than 50 billion CFU of probiotics)

[0233] Group B (n=20): Junerqing® Amoxicillin Clavulanate Potassium Dispersible Tablets + Composition of Comparative Example 1 (prepared according to the formulation of Comparative Example 1, 3g / bag, each bag contains no less than 50 billion CFU of probiotics)

[0234] Group C (n=20): Junerqing® Amoxicillin Clavulanate Potassium Dispersible Tablets.

[0235] 1.2.2 Administration and Testing

[0236] Participants were given one sachet of probiotics three times a day, either directly or mixed with warm water or milk (not exceeding 37°C) for seven days. Adverse reactions (incidence, severity, and duration) were monitored daily using a questionnaire. Fecal samples were collected on days 0, 3, and 7 for 16S rDNA sequencing to analyze gut microbiota diversity (Alpha / Beta diversity) and changes in species composition.

[0237] 2. Results

[0238] 2.1 Overall Usage

[0239] Table 9 Results of administration in each group

[0240]

[0241] As shown in Table 9, Group A had the lowest incidence of adverse reactions (20%) and the shortest duration of symptoms (<1 day). The recovery time of the primary disease in Group A was significantly better than that in other groups.

[0242] 2.2 Intestinal flora analysis

[0243] 2.2.1 Alpha Diversity

[0244] Table 10 Alpha diversity index of each group of samples

[0245]

[0246] As shown in the table above, for Observed OTUs: the bacterial community richness in group C decreased significantly (Δ=84.25), while the decrease in group A was smaller (Δ=6.33 / 10.62). Shannon index: the diversity in group A remained stable (Δ=0.01), while that in group C decreased significantly (Δ=0.32 / 0.35).

[0247] 2.2.2 Phylum-level species composition

[0248] See Figure 3 , Figure 4 Bacteroidetes / Firmwallis (F / B ratio): The F / B ratio in group A decreased from 1.22 to 0.67 (inflammation relief), while it remained at 1.09 in group C (residual inflammation); Proteobacteria (pro-inflammatory bacteria): The relative abundance in group A increased (5.54%→12.86%), which may be related to the clearance of pathogenic bacteria by antibiotics.

[0249] 2.2.3 Changes in key bacterial communities at the genus level

[0250]

[0251] Table 11 Composition of Major Fungal Genus in Each Group

[0252] As shown in Table 11, the abundance of beneficial bacteria (Bifidobacterium 4.42%→4.88%) and Bacteroides 18.19%→21.44%) increased in group A. The abundance of harmful bacteria (Aristobacterium diastereoides 4.23%→0.82%) decreased significantly in group A.

[0253] In summary, the probiotic composition of this invention can reduce the incidence of adverse reactions, shorten the duration of symptoms, accelerate the recovery of the primary disease, maintain the diversity of intestinal flora, increase the abundance of beneficial bacteria such as Bifidobacteria, and reduce the F / B ratio (inflammation relief), providing an effective solution for the precise intervention of antibiotic-related intestinal side effects.

Claims

1. A probiotic and prebiotic composition, characterized in that, It consists of the following components in parts by weight: 5-20 servings of probiotics 10-50 parts by weight of resistant dextrin 10-50 parts by weight of xylooligosaccharides 3-10 parts by weight of fructooligosaccharides 1-5 parts by weight of stachyose 5-10 parts by weight of erythritol 5-10 parts by weight of cranberry powder; The probiotic strains, by weight, are as follows: Bifidobacterium animalis subsp. lactis BLa80 10%-20% Pediococcus acidilactici CCFM7902 5%-15% Lactobacillus acidophilus LA85 5%-15% Lactobacillus plantarum N13 10%-20% Lactobacillus paracasei LC86 10%-20% Bifidobacterium bifidum BBi32 5%-15% Lactobacillus rhamnosus LRa05 10%-20% Lactobacillus casei LC89 5%-15%.

2. A method for preparing the probiotic and prebiotic composition as described in claim 1, characterized in that, The process includes the following steps: (1) preparing erythritol into an adhesive solution; (2) mixing resistant dextrin and xylooligosaccharide, spraying the adhesive solution prepared in step (1) into granulation, sieving and mixing it with stachyose, fructooligosaccharide, cranberry powder and probiotic powder to obtain the probiotic and prebiotic composition.

3. The method according to claim 2, characterized in that, In step (1), erythritol is prepared into a binder solution with a concentration of 10-50% (w / v); in step (2), the particle size is controlled to be 0.3-1.0 mm, the air inlet temperature during the granulation process is 50-70℃, the atomization pressure is 0.1-0.3 MPa, and the material fluidization velocity is 1.5-3.0 m / s.

4. The use of the probiotic and prebiotic composition according to claim 1 in the preparation of antibiotic adjuvants.

5. The application according to claim 4, characterized in that, The antibiotic adjuvant composition is used to relieve antibiotic-associated diarrhea.

6. The application according to claim 4, characterized in that, The antibiotic adjuvant is used in combination with antibiotics for the treatment of respiratory infections.

7. The application according to claim 4, characterized in that, The antibiotic adjuvant is used in combination with antibiotics to treat urinary tract infections.

8. An antibiotic adjuvant preparation, characterized in that, It comprises the probiotic and prebiotic composition of claim 1.

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